Redox active or thiol reactive? Optimization of rapid screens to identify less evident nuisance compounds. Issue 6 (June 2022)

• Record Type: Journal Article
• Authors: Proj, Matic; Knez, Damijan; Sosič, Izidor; Gobec, Stanislav
• Is Part Of: Drug discovery today. Volume 27:Issue 6(2022)
• Page Start: 1733
• Page End: 1742
• Publication Date: 2022-06
• Subjects: Redox -- Covalent -- Screening -- Hits -- PAINS -- Assays; ALARM NMR a La assay to detect reactive molecules by nuclear magnetic resonance -- Cys cysteine -- DPPH 2, 2-diphenyl-1-picrylhydrazyl -- DTNB Ellman's Reagent (5, 5-dithio-bis-(2-nitrobenzoic acid)) -- DTT dithiothreitol -- GSH reduced glutathione -- H2DCFDA 2', 7'-dichlorodihydrofluorescein diacetate -- HPLC high-performance liquid chromatography -- HRMS high-resolution mass spectrometry -- HRP-PR horseradish peroxidase-phenol red -- HTS high-throughput screening -- LC-MS liquid chromatography−mass spectrometry -- PAINS pan-assay interference compounds -- RCC redox cycling compound -- REOS rapid elimination of swill -- ROS reactive oxygen species -- TCEP tris(2-carboxyethyl)phosphine -- TNB2– 5-mercapto-2-nitrobenzoic acid; Drugs -- Design -- Periodicals; Drugs -- Research -- Periodicals; 615.1
• DOI: 10.1016/j.drudis.2022.03.008
• Languages: English
• ISSNs: 1359-6446

Abstract

Graphical abstract: Highlights: Redox active and thiol reactive compounds can lead to false positive screening hits. Optimization of four different high-throughput assays. Guidelines for detection of redox activity and thiol reactivity. New promiscuous compounds among bioactive compounds and chemical probes identified. Abstract: Compounds that exhibit assay interference or undesirable mechanisms of bioactivity are routinely encountered in assays at various stages of drug discovery. We observed that assays for the investigation of thiol-reactive and redox-active compounds have not been collected in a comprehensive review. Here, we review these assays and subject them to experimental optimization to improve their reliability. We demonstrate the usefulness of our assay cascade by assaying a library of bioactive compounds, chemical probes, and a set of approved drugs. These high-throughput assays should complement the array of wet-lab and in silico assays during the initial stages of hit discovery campaigns to pursue only hit compounds with tractable mechanisms of action.